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TTD Medium (DSMZ Medium 480b)

[所属分类:培养基配方] [发布时间:2021-9-29] [发布人:网站管理员2] [阅读次数:] [返回]
TTD Medium
(DSMZ Medium 480b)

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

NaCl (marine salts) ..................................................................... 25.0g
Sulfur, powder............................................................................. 10.0g
Casitone ........................................................................................ 5.0g
NH4Cl ......................................................................................... 0.33g
CaCl2·2H2O................................................................................. 0.33g
MgCl2·6H2O................................................................................ 0.33g
KCl.............................................................................................. 0.33g
KH2PO4....................................................................................... 0.33g
Resazurin ...................................................................................1.0mg
Na2S·9H2O solution .................................................................10.0mL
Vitamin solution.......................................................................10.0mL
Trace elements solution SL-10 ..................................................1.0mL
pH 6.9 ± 0.2 at 25°C
Trace Elements Solution SL-10:
Composition per liter:
FeCl2·4H2O................................................................................... 1.5g
CoCl2·6H2O ........................................................................... 190.0mg
MnCl2·4H2O .......................................................................... 100.0mg
ZnCl2........................................................................................ 70.0mg
Na2MoO4·2H2O ....................................................................... 36.0mg
NiCl2·6H2O.............................................................................. 24.0mg
H3BO3 ........................................................................................ 6.0mg
CuCl2·2H2O ............................................................................... 2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl2·4H2O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor
oughly. Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–
121°C.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................ 10.0mg
Thiamine-HCl·2H2O.................................................................. 5.0mg
Riboflavin .................................................................................. 5.0mg
Nicotinic acid............................................................................. 5.0mg
D-Ca-pantothenate ..................................................................... 5.0mg
p-Aminobenzoic acid................................................................. 5.0mg
Lipoic acid ................................................................................. 5.0mg
Biotin ......................................................................................... 2.0mg
Folic acid ................................................................................... 2.0mg
Vitamin B12 ................................................................................ 0.1mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Sparge
with 80% H2 + 20% CO2. Filter sterilize.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O.................................................................................... 0.5g
Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis
tilled/deionized water and bring volume to 10.0mL. Sparge with N2.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an
aerobically.
Preparation of Medium: Prepare and dispense medium under an
oxygen-free 80% N2 + 20% CO2 gas mixture. Add components, except
vitamin solution and Na2S·9H2O solution, to 980.0mL distilled/deion
ized water. Mix thoroughly. Sparge with 80% N2 + 20% CO2. Adjust
pH to 5.9 with concentrated NaOH. Sterilize medium by heating for 1
hr at 90°C–100°C on 3 subsequent days. Sparge with 80% N2 + 20%
CO2. Before use, aseptically and anaerobically add 10.0mL sterile vi
tamin solution and 10.0mL sterile Na2S·9H2O solution. Mix thorough
ly. Aseptically and anaerobically distribute into sterile tubes or flasks.
Use: For the cultivation of Thermococcus stetteri.
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